AIMTo elucidate the effect of sphingosine kinase (SPK) on the hepatocyte growth factor (HGF)-induced migration of endothelial cells.

METHODSWe constructed recombinant adenoviral vectors, which contain SPK gene and its mutant respectively. These adenoviral vectors were packaged and amplified in 293 cells. And intracellular SPK activity was assayed via measurement of [32]P radioisotope labeled S1P; the effect of SPK activation on HGF-induced migration of endothelial cell was observed by Transwell technique.

RESULTSAdenoviral mediated expression of SPK gene increased in ECV 304 cells intracellular SPK activity, which in turn enhanced the HGF-induced migration. Whereas these activities were blocked by the dominant negative SPK gene.

CONCLUSIONThese findings show that SPK activation plays important roles in the regulation of HGF-induced migration of endothelial cells.

Adenoviridae ; metabolism ; Cell Line ; Cell Movement ; drug effects ; Endothelial Cells ; cytology ; Hepatocyte Growth Factor ; pharmacology ; Humans ; Phosphotransferases (Alcohol Group Acceptor) ; genetics ; metabolism ; Signal Transduction